Gnao1 acts as a gatekeeper to alleviate neuropathic pain by silencing pro-nociceptive signaling cascades.

IF 8.7 1区医学 Q1 CLINICAL NEUROLOGY

Journal of Headache and Pain Pub Date : 2026-06-23 DOI:10.1186/s10194-026-02436-6

Jieyi Cai, Hui Li, Lü Chen, Ling Luo, Xin Wu, Min Yuan, Maolin Han, Shirong Wen, Changlai Zhu, Yun Gu

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引用次数: 0

Abstract

Background: Neuropathic pain is a debilitating condition with limited effective treatments, highlighting an urgent need for non-opioid analgesic targets. Gαo, encoded by the Gnao1 gene, is an abundant G protein in the nervous system and couples to multiple inhibitory GPCRs, but its role in chronic pain remains poorly understood.

Methods: Spatial transcriptomics and bulk RNA-sequencing were performed on the spinal cord and dorsal root ganglia (DRG) of mice following spared nerve injury (SNI). Adeno-associated virus (AAV)-mediated Gnao1 overexpression or knockdown was performed in the spinal cord or DRG, followed by pain behavioral testing using von Frey filaments, acetone evaporation test, and mechanical conflict avoidance (MCA) assay. Microglial activation, neuronal injury, and signaling pathways were examined by immunohistochemistry (IHC), Western blotting (WB), and calcium imaging. In vitro studies used BV2 microglia and primary DRG neurons with lentiviral Gnao1 overexpression and LPS or db-cAMP stimulation.

Results: Spatial transcriptomics revealed that Gnao1 is enriched in the spinal dorsal horn (SDH) and significantly downregulated after SNI. Bulk RNA-seq confirmed Gnao1 downregulation in pain-processing regions (DRG and SDH). IHC showed that its cellular localization shifted from neurons to glial cells. Overexpression of Gnao1 in the spinal cord or DRG markedly alleviated SNI-induced mechanical allodynia, cold hypersensitivity, and pain-related avoidance behaviors. Mechanistically, transcriptomic profiling revealed that Gnao1 overexpression broadly suppressed pro-inflammatory and pro-nociceptive pathways. Experimental validation confirmed that Gnao1 overexpression reduced p38, ERK, and NF-κB phosphorylation, and decreased microglial activation. In primary DRG neurons, Gnao1 overexpression also lowered cAMP levels and suppressed db-cAMP-evoked Ca²⁺ influx.

Conclusion: Gnao1 acts as a gatekeeper negative regulator of neuropathic pain by silencing distinct pro-nociceptive signaling cascades, and highlights its potential as a non-opioid analgesic target.

Clinical trial number: Not applicable.

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Gnao1作为一个看门人，通过沉默前伤害性信号级联来减轻神经性疼痛。

背景：神经性疼痛是一种衰弱性疾病，有效治疗有限，迫切需要非阿片类镇痛靶点。由Gnao1基因编码的Gαo是神经系统中丰富的G蛋白，与多种抑制gpcr偶联在一起，但其在慢性疼痛中的作用尚不清楚。方法：对脊髓和背根神经节（DRG）进行空间转录组学和大量rna测序。在脊髓或DRG中进行腺相关病毒（AAV）介导的Gnao1过表达或敲低，随后进行von Frey细丝疼痛行为测试、丙酮蒸发试验和机械冲突避免（MCA）试验。通过免疫组化（IHC）、Western blotting （WB）和钙显像检查小胶质细胞活化、神经元损伤和信号通路。体外研究使用慢病毒Gnao1过表达和LPS或db-cAMP刺激的BV2小胶质细胞和原代DRG神经元。结果：空间转录组学显示Gnao1在脊髓背角（SDH）富集，SNI处理后显著下调。Bulk RNA-seq证实Gnao1在疼痛加工区（DRG和SDH）下调。免疫组化显示其细胞定位由神经元向胶质细胞转移。在脊髓或DRG中过表达Gnao1可显著缓解sni诱导的机械异常性痛、冷超敏反应和疼痛相关回避行为。在机制上，转录组学分析显示Gnao1过表达广泛抑制促炎症和促伤害通路。实验验证证实，Gnao1过表达可降低p38、ERK和NF-κB磷酸化，降低小胶质细胞活化。在原代DRG神经元中，Gnao1过表达也会降低cAMP水平，抑制db-cAMP诱发的Ca2+内流。结论：Gnao1通过沉默不同的促伤害性信号级联反应，在神经性疼痛中发挥负调控作用，并突出了其作为非阿片类镇痛靶点的潜力。临床试验号：不适用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Headache and Pain 医学-临床神经学

CiteScore

11.80

自引率

13.50%

发文量

143

审稿时长

6-12 weeks

期刊介绍： The Journal of Headache and Pain, a peer-reviewed open-access journal published under the BMC brand, a part of Springer Nature, is dedicated to researchers engaged in all facets of headache and related pain syndromes. It encompasses epidemiology, public health, basic science, translational medicine, clinical trials, and real-world data. With a multidisciplinary approach, The Journal of Headache and Pain addresses headache medicine and related pain syndromes across all medical disciplines. It particularly encourages submissions in clinical, translational, and basic science fields, focusing on pain management, genetics, neurology, and internal medicine. The journal publishes research articles, reviews, letters to the Editor, as well as consensus articles and guidelines, aimed at promoting best practices in managing patients with headaches and related pain.